CN107106605A - Automation, multi-functional, engineering heart tissue culture and test biology reactor assembly - Google Patents
Automation, multi-functional, engineering heart tissue culture and test biology reactor assembly Download PDFInfo
- Publication number
- CN107106605A CN107106605A CN201580054406.0A CN201580054406A CN107106605A CN 107106605 A CN107106605 A CN 107106605A CN 201580054406 A CN201580054406 A CN 201580054406A CN 107106605 A CN107106605 A CN 107106605A
- Authority
- CN
- China
- Prior art keywords
- post
- tissue
- bioreactor system
- optical fiber
- culture hole
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/12—Well or multiwell plates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/22—Transparent or translucent parts
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/50—Means for positioning or orientating the apparatus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M31/00—Means for providing, directing, scattering or concentrating light
- C12M31/10—Means for providing, directing, scattering or concentrating light by light emitting elements located inside the reactor, e.g. LED or OLED
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Clinical Laboratory Science (AREA)
- Computer Hardware Design (AREA)
- Molecular Biology (AREA)
- Mechanical Engineering (AREA)
- Physics & Mathematics (AREA)
- Electromagnetism (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Improved tissue engineered biological reactor and test platform are devised, it is integrated with a variety of tests and stimulatory function.The system allows mechanically and electrically stimulating, medium is irrigated, and convergent force automatic monitoring and it is extracellular it is electroactive parallel under multiple strip of tissue growth.The system is designed to inexpensive and expansible, with the high content for providing the engineering musculature performance to being measured usually using muscle bath system, biological true to nature, nondestructive testing.
Description
Governmental support
The present invention is to be made on the basis of the UL RR029887 issued by NIH under governmental support.Government is in this hair
Possess some rights in bright.
The cross reference of related application
Present patent application based on and require that the U.S. of Serial No. 62/046,569 submitted for 5th in September in 2014 is interim
The priority of patent application, entire contents are incorporated herein in the way of integrally quoting herein.
Technical field
The present invention relates to for growing tissue's (for example, multiple heart tissue bars) under defined electro photoluminescence and mechanical stimulus
Bioreactor system, more particularly, to will measure and control function is integrated into improved life in tissue culture system
Thing reactor assembly, to allow to carry out automatic, measurement in real time with the tissue in the case where that need not contact.
Background technology
Many work have been carried out in recent years and are engineered heart tissue to develop the three-dimensional (3D) from multiple sources, it is main
The heart cell of rat myocardial cell that will be from separation and derived from human embryonic stem, and the multipotential stem cell induced now come
Source.These engineering tissues have been produced for a variety of purposes in a variety of forms, for example, develop for suffering from myocardial infarction (MI)
Patient damage heart tissue paster, and as the testing in vitro for screening potential therapeutic agent, toxotest
Platform, and it is used as the substitute of the animal model for studying health and illing tissue.
The common manifestation of latter object is created with electroactive and contraction activity engineering tissue bar.By these productions
Raw power is measured by various technologies and provides the functional performance characteristic of the tissue.One in these force measuring methods
It is a little to include optically tracked dimethyl silicone polymer (PDMS) contilever platform with post deflection, and use some sensor skills
The standard muscle bath force test device (Turnbull et al., 2013) of art.In the former, utilize what is separated at distance about 1cm
Two flexible columns manufacture small PDMS casting, and fibrin/collagen/3D engineering groups based on Matrigel are grown in-between
Knit bar.After culture a period of time, the tissue will produce the convergent force for causing post to bend.For optical tracking because bending is led
Cause the video platform of the displacement of the end of the post, can use for cantilever beam bending equations non-invasively and in real time
The convergent force that the estimation tissue is produced.
These systems have many with being used to measure the muscle bath system of the heart separated and skeletal muscle tissue bar in decades
It is different.One is that the tic power that PDMS is measured is anisometric, and this is opposite with muscle bath system.This may cause the tissue
Shrinkage is underestimated, and this is due to the fact that the maximum, force of generation reduces with the shortening and mechanical relief of tissue.In addition,
Different from the bath setting of standard muscle, the tissue can not be fully stretched to its optimum length, and which also limits maximum, force production
It is raw and make the tic power from different tissues relatively become complicated.To natural before attempting to use muscle bath system relatively
When organizing the data of many experiments, these differences are considered PDMS shortcoming.
Similarly, muscle bath is also restricted, including high cost, low yield, ambient non-sterile and causes due to clamping
Tissue damage.Therefore, all there is limitation in both systems, above-mentioned related to traditional design it is therefore desirable to provide that can overcome
Limitation and the bioreactor system of defect.
The content of the invention
Improved tissue engineered biological reactor and test platform are devised, it is integrated with a variety of tests and stimulates work(
Energy.The system allows mechanically and electrically stimulating, medium is irrigated, and automatic monitoring convergent force and it is extracellular it is electroactive it is parallel under
The growth of multiple strip of tissue.The system is designed to inexpensive and expansible, is surveyed with providing usually using muscle bath system
The high content of the engineering musculature performance of amount, biological true to nature, nondestructive test.
Brief description of the drawings
Fig. 1 is top and the side perspective view of bioreactor system according to an embodiment of the invention；
Fig. 2 is its sectional view；
Fig. 3 is its top view；
Fig. 4 is the perspective view of the bioreactor system；
Fig. 5 is the sectional view of the optical fiber cylinder as fiber tension sensor；
Fig. 6 is the top view of many tissue casting moulds；
Fig. 7 is the curve map of the displacement vs tension force for the parameter listed in table 1, it was demonstrated that nearly 2 times of sophisticated defection signal are put
To increase to being applied greatly (based on luminous point compared with the actual fiber tip displacement at the position in the power of organization applied)
Power sensitiveness；
Fig. 8 is the sectional view for the position sensitive detectors (PSD) being used together with Fig. 5 optical fiber cylinder；
Fig. 9 is the curve map for normalizing the deflection output that differential voltage (NDV) vs carrys out self-calibration routines, is shown excellent
Linearly；With
Figure 10 is the display screen of software, and it is configured with the various parts for controlling the bioreactor system and measurement
And record information.
Embodiment
According to the present invention there is provided improved bioreactor system, the bioreactor system is integrated with the system
Feature simultaneously enhances function, while consistently keeping tissue (such as strip of tissue) in optimal incubator environment.This need by
It is all measurement and control function be integrated into the tissue culture system, with allow it is automatic, it is real-time measurement without with tissue
Contact is there is provided improved Data Collection, convenience and to the tissue and the control of experiment.
As discussed herein, allow to grow according to the construction of the bioreactor system of the present invention and monitor one or many
Individual tissue samples (such as heart tissue bar (cardiac tissue strips)).The bioreactor system is also designed
Into realizing following target following detailed description of (it is non-limiting list)：
1. Various Tissues can be grown in the medium under controlled electrically and mechanically stimulation；
2. can in automatic measurement incubator the tissue passive and active convergent force, the length of tissue can be adjusted simultaneously
Degree and tension force and pacing frequency；
3. the action potential produced during shrinking by the tissue can be measured；With
4. built-in fluid communication port can be used easily to irrigate culture medium and test agent in culture hole (bath).
Fig. 1-4 shows the bioreactor system 100 according to one exemplary embodiment of the present invention.Fig. 1 is described
The top of bioreactor system 100 and side perspective view；Fig. 2 is the sectional view of the bioreactor system 100；Fig. 3 is
The top view of the bioreactor system 100；Fig. 4 is the perspective view of the bioreactor system 100.
As illustrated, the bioreactor system 100 can be placed at support surface (such as desk or length
Stool or incubator shelf etc.) on stand-alone assembly (unit) form.As described herein, the system 100 is Power Component, because
This is operatively coupled to power supply, such as power outlet, battery, combinations thereof.The bioreactor system 100 is included
Bottom support 110, it supports the electronic device of various workpieces and bioreactor system 100.The bottom support
110 can be the forms such as substrate.In an illustrated embodiment, the bottom support 110 includes the first support member 112, and it leads to
Multiple supporting legs 116 (it can be arranged in corner) are crossed to raise relative to the second support member 114.The support member 112,114 can
In the form of being plane tabular substrate arranged parallel to each other.Space between two support members 112,114 allows cable, visual
Standby wait of makeup route between them.
It should be appreciated that the bottom support 110 can be single structure form and/or can using be suitable for install
Any amount of multi-form of the part of the system 100.Or, the bioreactor system 100 need not necessarily be mounted on bottom
On portion's support member 110, but separate unit that is transportable and may be provided in support surface (such as desk or platform).
The bioreactor system 100 also includes the pedestal 120 with inner space.Therefore, the pedestal 120 is bag
The hollow structure of at least part containing other parts as discussed herein.The pedestal 120 can be formed as with any number
The different shape of amount, including but not limited to square or rectangular shape.Therefore, the pedestal 120 is by bottom plate and limits in hollow
Multiple side walls 122 in portion are limited.In the case of square base 120, the pedestal 120 includes four side walls 122.Therefore, institute
Unlimited top can be had by stating pedestal 120, can be close to hollow inside by the unlimited top.
As illustrated, the pedestal 120 can be arranged on the first support member 112 of bottom support 110.The pedestal
120 can be fixedly secured on the first support member 112 or can be simply resting on the first support member 112.
The bioreactor system 100 is also comprising the culture hole (bath) being arranged in the hollow inside of pedestal 120
130.Position and the construction of the culture hole (culture well) 130 are described below in detail.In general, the culture hole
130 represent engineering heart tissue grows wherein, and also tested in the case of this bioreactor system 100
Hole.Therefore, the culture hole 130 can be set along the bottom plate of pedestal 120.
As best seen in figure 2, the bottom (bottom plate) of the culture hole 130 can by such as slide transparent configuration
(printing opacity) 135 is limited.The size (such as height) of the culture hole 130 is by the High definition of pedestal 120.
The bioreactor system 100 can also include the lid 140 coordinated with pedestal 120.Therefore the lid 140 is closed
The unlimited top of pedestal 120.Therefore the lid 140 can be the planar structure for the open-top for being positioned across pedestal 120, be used for
The hollow inside of closed base 120.The lid 140 can have the flat top surface 142 for installing equipment.The lid 140 can be from
Pedestal 120 is removed.As described herein, during the assembling of system 100, the lid 140 is reduced on pedestal 120 with closed base
120.It can use any amount of different positioning, coupling and/or locking mechanism that lid 140 is fixedly attached into pedestal
120。
The bioreactor system 100 also includes force sensor module (force snesor/Force sensor) 300.It is described
Force sensor module 300 is configured with the passive and active convergent force for the tissue that measurement grows in culture hole 130, and can be simultaneously
Adjust the length and tension force of tissue.
Force sensor module 300 according to an embodiment of the invention includes two critical pieces, i.e. first sensor
Part 310 and second sensor part 350.The first sensor part 310 is configured to its part and extends to culture hole 130
In, and the second sensor part 350 is arranged on the outside of culture hole 130.
The force sensor module 300 can be installed on the support member 305 separated with lid 140 and pedestal 120.The branch
Support member 305 be designed to by the fixed position of force sensor module 300 and be maintained at relative to culture hole 130 fixed position (and
Therefore relative to lid 140 and pedestal 120).The support member 305 can be the form of planar substrates (as shown in the figure), when lid 140
When being coupled to pedestal 120, the planar substrates are located at the top of lid 140.For example, the support member 305 can be with elevated side
Formula is installed to support member 112, and especially, support member 305 can be raised by multiple supporting legs 307 relative to support member 112.
The supporting leg 307 is contacted and leaned against on support member 112, more specifically, supporting leg 307 is fixedly attached to support member 112, to incite somebody to action
Component 300 is locked in the appropriate location on lid 140.Therefore, the component 300 can be dismountable and can be from support member
110 remove, while being regularly locked in relative to support member 110 and therefore relative to the setting of pedestal 120 and lid 140
Position.
The support member 305 is considered the pedestal of component 300, and allows the equipment of force sensor module 300
It is mounted thereto.The support member 305 extends laterally across the width of pedestal 120 and lid 140.The support member (pedestal) 305
With top surface 309.The support member 305 can include the electronic device associated with component 300.For example, peace can be provided
Be attached to the printed circuit board (PCB) 312 of the support member 305, and provide for first sensor part 310, and for power supply and
Installation of other signals to the connector of outside control, amplification and/or data acquisition equipment.Although the circuit board 312 is shown
Go out for relative to the right angle setting of support member 305, but circuit board 312 is not limited to be in this orientation, but it can be arranged to
It is oriented, for example, be horizontally mounted.
The first sensor part 310 is the form of illuminated component, more specifically including at least one light source, and it is configured
Light is guided for the axis in predetermined direction along restriction.In a change case, the first sensor part 310 includes vertical
The LED/light source of installation so that the light launched from LED/light source is downward along the axis perpendicular to lid 140 by optical fiber tissue post 340
Point to.
In the embodiment shown, the first sensor part 310 includes multiple light sources, more specifically, in a change
Multiple LED/light sources are included in example.For example, in the embodiment shown, exist and be installed to the circuit board 312 (and operationally
Be connected to processor 312) three LED/light sources 310.Three LED/light sources 310 can pacify along identical axis linearity
Dress.The axis for the light launched from three LED/light sources 310 is parallel to each other and in relation spaced apart.Therefore, each
Being used for comprising their own of LED/light source 310 launches the LED of light, but can also be coupled to common light source.
According to the present invention, each LED/light source 310 includes optical fiber LED emitter 330 and associated optical fiber tissue post 340,
The optical fiber tissue post 340 is operatively coupled to optical fiber LED emitter 330, is projected with allowing light through optical fiber tissue post 340.
Therefore, the optical fiber tissue post 340 is formed by optical fiber.It is well known that optical fiber is by extruding glass (silica) or coating modeling
Expect flexible, the transparent optical fiber being made, and optical fiber is transmitted as waveguide or light pipe between the two ends of optical fiber.Therefore, it is described
Optical fiber LED emitter 330 is used to the light from LED/light source (bulb) being transmitted into optical fiber, and the optical fiber is light in this case
The form of fibre tissue post 340.This alignment between the transmitter 330 and optical fiber tissue post 340 causes light in culture hole
Guided downwards in 130, more specifically, light is axially guided into second sensor part 350, as discussed herein, second biography
The detection light of inductor components 350.
The first sensor component 310 extends in support member 305 and both lower sections of lid 140, and therefore, the two structures are matched somebody with somebody
It is set to each optical fiber tissue post 340 of receiving.For example, the lid 140 can have the opening that optical fiber tissue post 340 is passed through.It is described
Optical fiber tissue post 340 is axially aligned and in the culture hole 140 so that they be typically normal to the bottom plate of culture hole 140 and
It is parallel to each other and axially align.The opening formed in the lid 140 can have containment member, such as rubber washer, O shapes
Ring etc..
The bioreactor system 100 can be assembled with any amount of different modes, and order or component can be with
Changed according to the construction of part.For example, the support member 305 can be installed after lid 140 is installed to pedestal 120, because
Support member 305 extends on lid 140 and crosses lid 140.The circuit board 312 associated with the optical fiber tissue post 340 allow for
Optical fiber LED emitter 330, and pacing electrode 600, metal structure post 400 and other local or outside its containing circuit board
It, which is connected, provides power supply.
The second sensor component 350 can be provided in below the dianegative 135 (slide) of culture hole 130
The form of sensor (for example, position sensitive detectors (PSD)), and it is configured with first sensor component 310 (post 340) detection
It is mobile.The operation of the second sensor part 350 is described below in detail.
The bioreactor system 100 is also comprising one or more moveable tissue posts 400, the tissue post 400
It is arranged in culture hole 130 and is a part for tissue column assembly 401.According to the present invention, there are multiple moveable tissues
Post 400 (for example, being three in the embodiment shown), and movably the quantity of tissue post 400 is equal to optical fiber tissue post 340
Quantity.The moveable tissue post 400 is arranged so that them towards optical fiber tissue post 340 and spaced away, therefore,
One group of post includes an optical fiber tissue post 340 and a moveable tissue post 400.In an illustrated embodiment, thus exist
Three groups of posts, because in the presence of three optical fiber tissue posts 340 and three moveable tissue posts 400.Every group of post 340,400 is used for
The component of growing tissue in-between, as described below.
The moveable tissue post 400 is operatively coupled to controllably move the mechanism 410 of tissue post 400, more
Specifically, with controlled linear mode movement tissue post 400.These posts are rigid and conductive, so as not to bend and use
Make the sensing electrode of the electrical activity of tissue.The mechanism 410 can be the form of actuator, such as linear actuators, and its is online
Property controllably moves tissue post 400 on direction.The linear actuators include motor, such as stepper motor, it allows to removable
The accurate control of dynamic tissue post 400.
The tissue column assembly 401 is configured as being fixedly attached to the top surface of lid 140.When the mechanism 410 is line
Property actuator form when, mechanism 410 can be comprising being coupled to actuator base 412 and actuator module 414 on lid 140
(for example, linear movement module).The mechanism 410 includes what is controllably driven by the motor in actuator module 414
Drive shaft 416.The drive shaft 416 can be on the direction towards post 340,400 from the anterior to extension of actuator module 414
Stretch.
The tissue column assembly 401 also includes column base 420, and the column base 420 is coupled to linear slide block 430 and carries tissue
Post 400 so that the movement of column base 420 is directly translated as the movement of tissue post 400.The linear slide block 430 is slided without friction
Block, it allows column base 420 to be moved in a controlled manner on linear direction.The drive shaft 416 is even coupled to post by connecting rod 425
Seat 420, and the motion of drive shaft 416 is transferred to the column base 420 for carrying tissue post 400.Therefore, the column base 420 can be with
Including at least the cross member of the width extension across pedestal 120 and lid 140, and the sliding block represents the horizontal stroke from column base 420
The parallel arms spaced apart extended back to component.
The tissue column assembly 401 is also operatively connected to master controller (for example, processor), and it allows to be based on user
The instruction (input) of input controls and accurately moves tissue post 400.Therefore, the master controller and actuator module 414
(motor) communicates, and controls the driving of drive shaft 416, and therefore controls column base 420 and tissue post 400 on linear direction
Movement.Under the operation of the actuator module 414, tissue post 400 can be on the direction towards optical fiber tissue post 340
(so that reducing the spacing between post 340,400) or on the direction away from optical fiber tissue post 340 (so as to increase post 340,400
Between spacing) driven, or these movements any combinations to produce arbitrary linear stretch/shortening scheme.
In the embodiment shown, the moveable tissue post 400 is the form of tungsten post, thin, rigid, raw to provide
The compatible tissue installed part of thing, when being connected to the input of appropriate amplifier, it can also detect extracellular electrical conductivity.
When the lid 140 is assembled into pedestal 120, moveable tissue post 400 and optical fiber tissue port 340 are arranged on training
Support in hole 130, its remote (bottom) end is spaced apart with the bottom plate of culture hole 130.
As explained below, the optical fiber tissue post 340 is substantially flexible and is suspended in culture hole 130, because
This, when applying enough power, optical fiber tissue post 340 shows bending (degree of free end lateral displacement), and (tungsten) firmly
Tissue post 400 keeps straight without bending.
As shown in fig. 6, tissue casting mould 500 can be by being placed on the suitable non-stick enclosure material of the bottom of culture hole 130
For exampleManufacture, is aligned (referring to Fig. 6) by the protuberance 510 in the slit that is fitted on the either side in hole 130.Three
Individual dog bone hole 520 is located at the lower section (Fig. 2 and 3) of the post 340,400 so that tissue is around the formation of post 340,400.The mould
The end of tool is more deeper than the length thereof of mould, and there is provided the hole of the agarose filled with fusing.Once the agarose is cooled to
Firm hydrogel, post 340,400 is pushed into hole through agarose, and is organized in the top of agarose and is formed so that
Once removing tissue from mould, the end of post 340,400 just exposes.
According to the scheme formation tissue (example developed by Costa Lab using previous PDMS systems (Serrao etc., 2012)
Such as strip of tissue).In short, the ice-cold collagen-matrigel-cell mixtures of about 30 μ l are pipetted into each hole, then place
The lid so that tungsten post and optical fiber cylinder 400,340 are aligned with the relatively deep end bag portion of casting mould 500 and pierce through agarose, then
Cultivated 2 hours before culture medium is added.Carefully lift the lid 140 after 48 hrs with from casting mould 500
Tissue is removed, mould 500 is removed, electrode is easily reconnected using magnetic pin joint mouthful, and reinstall lid 140.Then it is accurate
Get the system ready and carry out normal operating and measurement.
Perfusion system can use port to implement, and the port is located in the side of the culture hole 130 and biological anti-
Outside the side for answering device system 100.This allows press-fit cock or similar connection equipment to connect to circulate or exchange culture with pipe
Base.
The bioreactor system 100 is computer implemented system, and can include one or more computing devices,
And in one aspect, the bioreactor has the structure communicated by data acquisition and control interface with outside computing system
Part.The computing device can be the forms such as personal computer, mobile device, tablet personal computer, workbench.Computing device includes use
In performing software code with one or more processors of the operation of control data processing unit, read-only storage (ROM), random
Memory (RAM) or any other suitable volatibility or non-volatile computer readable storage medium storing program for executing are accessed, it can be solid
It is fixed or moveable.The system 100 preferably include one or more network interfaces, with by communication network to from
Other computing device transmission and reception data.The network interface can be realize communication between any equipment any
Interface, and comprising (but being not limited to) modem, NIC (NIC), integrated network interface, radiofrequency launcher/
Receiver (for example, bluetooth, mobile phone, NFC), Satellite Communication Transmit device/receiver, infrared port, USB interface and/or use
In any other such interface for connecting the equipment and/or communication network (such as dedicated network and internet).It is such
Connection can include wired connection or wireless connection (for example, using the known standards of IEEE 802.11 in association area), still
It should be appreciated that network interface can essentially realize any interface to/from the communication of processor.
The computing device used within system 100 includes internal memory, and generally comprises storage device.It can include all
Such as hard disk drive, floppy disk, tape drive, CD-ROM or DVD drive, flash memory, CD-RW, rewritable magnetic
The storage device of band, cloud storage or some above-mentioned combinations, for store program codes, database and application code.
In some embodiments, internal memory and/or storage device can be accessed by the processor so that processor can receive and
Perform the instruction being stored on internal memory and/or memory.In addition, element includes one or more input equipments, such as keyboard, mouse
Mark, trace ball etc., and display.The display can be comprising screen or any other such display device, and it causes
The system can indicate or otherwise provide a user the feedback of the operation on system (100).As an example, display
Device can be digital display, such as LCD display, CRT, light-emitting diode display or skilled artisans will appreciate that it is other this
The two dimensional display of sample.As another example, user interface and display are desirably integrated into touch-screen display.Therefore, institute
State display to be additionally operable to show graphic user interface, it can show various data and provide inputs information comprising permission user
" list " of field.Touching the position of the touch-screen of the display corresponding to graphic user interface allows user to be handed over equipment
Mutually with input data, control function etc..Therefore, when the touch screen is touched, this change is conveyed to processor by interface, and
Setting can be changed or the information of user's input can be captured and stored in internal memory.
One or more software modules can be coded in the storage device and/or internal memory.The software module can
With including one or more software programs with the computer program code or instruction set performed within a processor or application.With
Can be with ability in the operation of execution system and method disclosed herein or such computer program code of aspect or instruction
Any combinations of one or more programming languages that field technique personnel are understood that are write.Described program code can be as independent
Software kit is performed completely on a computing device (for example, data processing equipment), partly on one device and part
Held in one or more remote computing devices (such as user calculating equipment), or completely in such remote computing device on ground
OK.In the latter case, the various computing devices (can include LAN by any kind of wired or wireless network
(LAN) or wide area network (WAN)) be connected to media server data processing unit, or can be the company that outer computer makes
Connect (for example, by Internet use ISP).It should be appreciated that in some illustrative embodiments, Ke Yitong
Cross network and download one or more software modules via the network interface from another equipment or system.For example, being stored in service
The program code in computer readable storage devices in device from server can download to memory by network.
Optical Fiber Force Sensor
As described herein, first and second sensor element 340,350 is used as Optical Fiber Force Sensor.The optical fiber
(first sensor part) 340 is hung vertically in culture hole 130, the terminal growth for being organized in optical fiber 340.The optical fiber
Organize post 340 that as the light guide of second sensor part 350, the cantilever for measuring tissue contracts, the second sensor were both used for again
The form of part 350 is position sensitive detectors (PSD) 350.The position sensitive detectors 350 are can be in sensor surface
(PSD determines light to upper one-dimensional or two-dimensionally measurement light spot position optic position sensor using photodiode surface resistance
The position (place) of point).In the current situation, the luminous point is produced by optical fiber LED emitter, and in position sensitive detectors
Optical fiber is passed through before luminous point is produced on 350.The PSD 350 is arranged on below the slide for the bottom plate to form culture hole 130, and
Allow to pass through (transmission) from the light that the open distal end of optical fiber tissue post 340 is launched.The PSD 350 is connected to transimpedance amplification
The low current produced by PSD 350 amplification is converted into measurable voltage by device, transimpedance amplifier configuration.
Following information is described in detail the mechanically performance of example fiber tissue post 340 and its how measured with power
Ability correlation (it should be appreciated that following size and characteristic are only exemplary, rather than limitation of the present invention)：
Fibre diameter：0.5mm (CK-20, Industrial Fiber-optics, Inc.)
Optical fiber E modulus：4GPa
The optical fiber property and its configuration determine its response to tension force and the resulting pole of photoelectricity two detected
Pipe signal (see Fig. 5).Fig. 5 is the sectional view of optical fiber tissue post 340 for showing to be draped down from lid 140, wherein optical fiber (post 340)
Top be more rigid part.As illustrated, the free distal end of the optical fiber tissue post 340 is with being arranged on second sensor
The slide of the top of part 350 is spaced apart.
It is assumed that tension force p occurs at a point on optical fiber, at fixing end x.Lateral displacement γ at x is：
Wherein I is the area inertia moment (area moment of inertia) for the cylinder ellbeam that radius is r：
As shown in figure 5, the deflection γ at point x represents to be attached at the optical fiber tissue post 340 and removable tissue post 400
Between tissue length change.This does not indicate displacement of the luminous point on the sensor (PSD) 350, and this also depends on optical fiber
340 length, the angle of bend of optical fiber 340, and optical fiber 340 end and the distance of sensor 350.
The deflection d of optical fiber end end is given by：
Beam end is given by point x relative to the angle, θ x of vertical line：
As light beam travel distance D to sensor (PSD 350), the angle produces the additional lateral displacement δ of light beam, under
Formula is provided：
δ=D tan θx
Formula 5
Total spot displacement Δ is given by：
Δ=d+ δ
Formula 6
Following parameter and it is worth for the system modelling：
Table 1：Tension sensor beam performance modeling parameter
| Parameter | Description | Numerical value | Unit |
| r | Fiber radius | 0.25 | mm |
| l | Fiber lengths of the pedestal to tip | 15 | mm |
| x | Distance of the pedestal to power of organization | 13 | mm |
| D | Distance of the fiber optic tip to fibre optical sensor | 4 | mm |
| E | The Young's modulus of optical fiber | 4.2 | GPa |
| p | Tissue is applied to the point power on optical fiber | 0-1000 | μN |
Fig. 7 is the curve map of the displacement vs tension force of the parameter provided in table 1.This two curves are shown due to tissue contracts
The fiber optics displacement and resulting spot displacement (that is, the Δ in above-mentioned formula) of (that is, the γ in formula l) at x.Note described
How force snesor design provides the increased deflection sensitivity to tissue contracts power (signal gain).
Optical Fiber Force Sensor-PSD Amplifier Designs
The position sensitive detectors (PSD) 350 include the amplifier of the sensitive zones with 3mm length.Sensor 350
It is lateral photodiode, the photodiode has two electrodes, and common cathode in the either end of anode.From the light
Fine light beam is received by PSD 350 anode region, and it serves as the distributing switch between two electrodes, based on close to any electricity
The luminous point distribution electric current of pole.Fig. 8 is the sectional view for the structure for showing the PSD 350.
The illustrative sensors 350 used are Hamamasu S8673 PSD.The LED/light source (optical fiber 340) can
To be commercially available trade name Industrial Fiber-Optics IF-E92B optical fiber.
The LED can have 470nm wavelength, and incident optical power on sensor 350 is estimated as 20 μ W.It is described
PSD 350 can have 0.2A/W sensitivity at 470nm, and this will produce 4 μ A total current Io.
It is given by by the electric current of PSD 350 each electrode：
For 3mm Lx, 0.05mm change x, and 4uA Io, about 67nA is turned to by the change of the electric current of each electrode.
Minimum detectable change desired for 0.005mm places, electric current is 6.7nA.The comprehensive assessment of detection range is 0.1mm,
The difference of generation 0 to 150nA between electrode.
Twin spans impedance amplifier, such as commercially available trade name ADA4000-2 amplifier, available for amplification from each electricity
The electric current of pole.The gain bandwidth product (GBP) of the amplifier is 5MHz, and input capacitance is 5pF.Initial plate will have 10pF in parallel to feed back
The 1Mohm of capacitor feedback resistor.The maximum magnitude of output of the PSD amplifiers in the configuration of its electric current has been tested
Try as from 0 to -7.7V.This is due to the fact that only negative current can occur by each electrode.Another suitable plate can
To be biased using the divider for the non-inverting input for being applied to operational amplifier, to provide+7 to -7V effective range.This will permit
Perhaps larger feedback resistor and increased Full-span output are used.In current design, each μ A of electrode 2 input is then defeated
Go out EoFor：
Eo=-Isignal*Rf+Vos=-2e-6*1e6+0V=-2V
Formula 8
For 150nA current swing, this corresponds to the 0.15V outputs from each electrode and swings that (and 0.3V's is net
Difference).This can be increased by changing foregoing circuit.
Raw voltage signals from each magnifying electrode electric current are by data acquisition instrument (such as National
Instruments DAQ) collection, and handled using appropriate software (such as LabVIEW).For a sensor from each
The voltage of electrode is by below equation, to obtain so-called " normalization differential voltage " (NDV)：
Then the normalization differential electrical of 0.15 volt of full scale swing (full-scale swing) can be calculated such as following formula
Pressure：
NDV=(2.15-1.85) * 10/ (2.15+2.85) NDV=.75NDV
Signal this means the system by generation from 0 to 0.75NDV, the scope for power is about 0 to 1000 μ N.
The NDV signals are calibrated to the displacement of optical fiber to measure power.Starting every time using the system to grow and
Before test organization, following (semi-automatic) calibration procedures are carried out.
It should be appreciated that what above-mentioned equipment and above-mentioned size and value and program were merely exemplary in itself, without limiting
The scope of the present invention.
According to the present invention, the bioreactor system 100 is arranged in incubator etc., and the incubator etc. is promoting to cultivate
Operated under the condition (" condition of culture ") of the tissue growth between post 340,400 in hole 130.
For the shell of power supply, the unit activity amplifier and PSD amplifiers hold together with bioreactor
It is contained in incubator, it is remote to pass the signal to that wherein Ethernet cable (or other communication cables) runs to DAQ from amplifier
Journey desktop computer.Or, it can implement wireless between various computer equipments of the present invention and bioreactor part
Communication.The M3-L of actuating module 414 implements to use USB cable.Flat cable be used for through the incubator preceding doorjamb or
Instrument port afterwards.
In building the bioreactor system 100 and performing above-mentioned calibration procedure, by three all fiber cuts
Into certain length and it is carefully positioned in their LED axles, and motor column base is normally supported the tungsten filament for tissue, the electricity
The cross bar extension that column seat is run below in optical fiber is replaced.This cross bar is in the height selected meticulously (to match described group
The height knitted) on side draw optical fiber, and LabVIEW programs record PSD output to the position simultaneously.Produce NDV
Than the linear graph of (versus) position, the slope of wherein fit line is the sensitivity in units of NDV/ μm.
Matlab scripts are used for geometry and displacement height based on the calibration value and the optical fiber and calculate the power
Sensitivity.It is given by μm/force sensitivity FSs of the NDV for the displacement sensitivity k of unit, in units of μ N/NDV formula is as follows：
In order to minimize evaluated error, corrector strip must apply displacement at the precise height that the tissue will grow,
And the method based on image should be used to measure the height of each optical fiber, with confirmatory measurement and consider at the rear place of tic power
Adjusted during reason analysis.If the height is different from the position where tissue, based on the formula (ginseng listed in previous section
See the 3rd to the 6th formula for example listed above), it is necessary to which NDV is converted to power by more complicated model.Representativeness calibration is bent
Line is shown in following Fig. 9.The high linearity in 100 microns of big ranges of deflection is exported the figure illustrates sensor.
Length and loading are controlled
The bioreactor system 100 of the present invention is designed to provide the length control of tissue with 1 micron of resolution ratio.
NewScale Technologies Piezoelectric Driving micro linear actuator provides the absolute position control of +/- .5 micrometer resolutions
System, with adjustable speed and acceleration.The motor is controlled by LabVIEW control softwares, and is communicated by USB.
The axle of the actuator is coupled to the tungsten tissue post 400 relative with Fibre Optical Sensor post 310.The tungsten post 400 passes through micro linear
Ball bearing sliding block is arranged on and is maintained in the bearing above culture dish, and motor pushing and the pulling bearing, in the same manner
All three mobile tissues are (referring to Fig. 3 and Fig. 4).It should be noted that realizing other linear actuators technologies (such as stepping electricity
The actuator of machine driving), to provide identical function.
The function provides the very big advantage more than PDMS and other systems controlled without length, for example, allow to tissue
Mechanical stimulus is carried out, it is determined that (cardiac specialist is known as L to optimum length physiologicallymax), and tested to measure contraction
Force-length relationship.In addition, this allows measurement actively and passively to organize rigidity, and also allow to load controllable rather than length
Controllable tested tissue agreement.
Electro photoluminescence and monitoring
The bioreactor system 100 also by for be organized into it is ripe and pace-making carbon electrode 600 combine electro photoluminescence,
And the amplifier of the extracellular electrical activity for monitoring tissue.The stimulating electrode 600 can be located at strip of tissue (in post
310th, grow between 400) either end parallel graphite cake form, to produce electric field in the whole length of tissue.
The electrode 600 can be connected by stainless steel tube connector with one or more embedded magnets, the insertion magnet upward sliding
Stainless steel engaging pin on lid 140.This helps easily to remove and reinsert electrode in experimentation, wherein described
Electrode 600 will be because electrolysis by-products are gathered and need to change and cleaning.By the steel pin 603 associated with the lid 140 in fact
Now with stimulating the electrical connection of distribution.
Impulse waveform is controlled using all LabVIEW as described in more detail controller/software.Configuration has can
Control the biphasic square wave pulse of duration, amplitude and frequency and sent from the simulation output port on data acquisition interface (DAQ)
To electrode 600.This provide simple, cheap and effective means, with the training period with experiment during tissue the whole longevity
Various time points during life stimulate and pacemaker tissue.
The present invention can use biopotential amplifier, particular according to one embodiment, can be based on Cornell University
Bruce Land design construct one group of bioelectric amplifier, such as his publication " Tools in 2001
Physiology Labs：An Inexpensive High-Performance Amplifier and Electrode for
Extracellular Recording " (Land etc., 2001) are summarized.The amplifier connection monopole measurement configuration, wherein
Tungsten filament is used as measuring electrode, and silver-silver chloride silk is arranged in the lower section on graphite cake opposite to provide common reference electrode.It should manage
Solution, what above-mentioned amplifier was substantially merely exemplary, and other amplifiers can be used.
Controller/software
As described herein, the bioreactor system 100 is a part for computer implemented system, and software is used for
Control the operation of the various parts of bioreactor system 100 and measurement and record data during test/experiment model.
For example, the LabVIEW programs developed provide real-time control during whole Therapy lasted to bioreactor
And data acquisition, it can be maintained 3 weeks or the longer time.The LabVIEW programs simultaneously monitor all three tissue power and
Electrical activity, and control the motor and electro photoluminescence.Interval data record and electrical pacing (electrical pacing) can be set
It is set to user-defined interval and duration generation.Have been carried out two experiment modules study power than length and power compared with
Fight frequency.These modules allow user to increase length or pacing frequency with defined increment and duration.The motor may be used also
To be set to vibrate between two points, stimulated with the cyclic stretching for providing tissue.
Figure 10 is the screenshot capture of an exemplary L abVIEW control program.
As simultaneously refer to the attached drawing is described in detail herein, the bioreactor system 100 is designed to realize following target：
Target 1- can grow Various Tissues in the medium under controlled electro photoluminescence and mechanical stimulus
First aim is the polysulfones sheet material that manufactures using CNC to realize, it includes culture hole 130 and supporting tissue and thorn
Swash component.Mechanical stimulus is realized (referring to second by the micro linear actuator 414 controlled with closed loop absolute position
Details in target).In a change case, the arm of the actuator is connected to the battle array of the tungsten post 400 of three 0.5mm diameters
Row, it supports three tissues (bar) abreast to grow.The relative side of the culture hole includes vertically arranged load measurement column (optical fiber cylinder
340) with another end of supporting tissue (referring to second target).It is graphite plate electrode on the outside of post array, with tissue
Whole length on electro photoluminescence is provided.Preferably, tissue growth in triple arrays to provide system 100 higher handle up
Amount.
Target 2- can in automatic measurement incubator the tissue passive and active convergent force, and being capable of regulation group simultaneously
The length and tension force and pacing frequency knitted
Second target is completed by optical fiber deflecting force sensor-based system.The plastic optical fiber of 500 micron diameters of standard
It is described to be organized in attachment and slight bending when it shrinks as hard but flexible post 340.Positioned at the end of optical fiber 340
The position sensing photoelectric detector (PSD) 350 of lower section receives the LED light for being transmitted through optical fiber 340, and detection fiber 340 from
By the displacement held.The photo detector signal is recorded and the formula based on bending cantilever beam is converted into displacement and tissue is produced
Power.Length of tissue is controlled by the way that the other end of tissue is attached into rigid tungsten filament (post 400), the rigid tungsten filament is connected to line
Property motion module 414, for example can be from NewScale Technologies, Victor, M3-L linear movement moulds commercially available NY
Block, the example modules are the piezoelectric linear actuators with absolute position control, 0.5 micrometer resolution and 6mm strokes.
Target 3- can measure the action potential produced during shrinking by the tissue
3rd target is entitled " the Tools for physiology labs published using Land et al. in 2001：
a inexpensive high-performance amplifier and electrode for extracellular
Amplifier Design in recording " (Land etc., 2001) paper is realized.The input of the amplifier is connected to branch
Support the tungsten post 400 of tissue, enabling cell is carried out to the action potential produced by strip of tissue (being grown between post 340,400)
Outer record.Suitable software (such as LabVIEW softwares) and other equipment (such as National Instruments data acquisitions
Plate (DAQ)), the Data Collection available for motor control, from all amplifiers, and provide boost pulse to electrode.
Target 4- can use easily perfusion culture medium and the test in culture hole (bath) of built-in fluid communication port
Reagent
4th target is to realize that tube connector can by the threaded port in the side of the culture hole 130 of bottom
With on the side of bioreactor system 100.Pipeline is routed to peristaltic pump to be used with appropriate speed from the port
Fresh culture (or containing soluble biochemical compound or for the culture medium for the medicament for measuring dose response) exchanges culture hole
In culture medium.Cock and the pipeline are in line placements, to allow the pipeline during structure observation and user and system interaction
Disconnect without leaking.
Therefore, the advantage of the bioreactor system 100 includes but is not limited to following：
A) the length control permission during the non-invasive monitoring of contractile function is (known in flesh in consistent baseline length
Lmax percentage is used as in meat physiology technics) middle relatively different tissues or different conditions, to allow length dependent
Just and standardization the comparison of convergent force.This is the standard practices in muscle physiological experiment, and has been applied to from culture ring
The engineering tissue removed in border carries out short-term test.But this damaging tissue, it can not be returned for subsequent culture and survey
Examination.The system allows integrated length to control and power is measured for testing for a long time, the sterile culture without destroying engineering tissue
Condition.Length of tissue is set to control motorization to allow comprehensive power-length testing agreement repeatedly and non-invasively to be performed, and
And several tissues are attached to the higher throughput test that identical motor allows the equipment economy and applied for screening.This
The row of a little motors can be coupled to allow tested tissue array extending.
B) the mobile permission newel post deflection of the light transmitted by optical fiber is monitored using photoelectric detector, so as to allow to shrink
Power in multiple tissues automatically, continuously while ground is monitored, and avoid due to big call data storage
The need for current system is limited to video surveillance or video acquisition.The temporal resolution of optical signal is also than most of video cameras
System is (generally<100fps) much higher (kilohertz range), to ensure the high-fidelity of the most swift nature even for the cycle of tic
Capture.Compared with the video camera or electronic force sensor (several thousand dollars) of replacement, our sensor-based system cost is low (several dollars),
This helps economically to expand to multiple tissues in single bioreactor platform.
C) it is still actual for intermittent inspection without being removed from incubator due to the optic visualization of the tissue
Need, the bioreactor system is arranged on below the supporting leg with inclined mirror.Therefore, camera can be placed on
Beside the bioreactor and focus on mirror, it provides the live image of engineering tissue.
D) electric stimulation electrode is placed in culture region is stimulated with the field for applying engineering tissue in the training period.The electricity
Pole and motor are electric synchronous so that pacing signal and response of twitching are coordinated in time.This is for ensuring stretch signal in flesh
Appropriate time during meat is twitched is vital.This idea forms referred to as cardiac resynchronization therapy (CRT)
The phase precise coordination of the timing on the basis of clinical technology, wherein electric pulse and cardiac cycle is so that the blood-pumping function of heart is maximum
Change.
E) because the carbon electrode is porous, they tend to that material is absorbed and accumulated from tissue-culture well (bath) environment
Material, which compromises their performance, and will be unfavorable for using in subsequent experiment.For the slow of the heart cell in culture
Property electro photoluminescence other business systems (for example, c-pace systems from Ionoptix) require their instrumentation lid have forever
The carbon electrode being attached long, " needing every 24-48 hours cleaning ", by using " being immersed in agitator in distilled water several days ", this is
Inconvenient and time-consuming, and final etching electronic circuit board component.In the present invention, detachable electrode using conductive magnet with
Electrode is fixed on position during use, with easily remove electrode be used for clean and rapidly with clean electrode replacement
They, and be firmly held in place electrode when operating the bioreactor.
F) in order to ensure light is launched from the end of the optical fiber, and in order to ensure tissue does not slide from end, it grows
In the sophisticated distance away from optical fiber connector post nearby (similarly on the tungsten end post of the other end of tissue).Therefore,
Special low-adhesion but permeable tissue through developing the unique combination using teflon and agarose aquogel are cast
Mould.Teflon tissue casting mould is placed on to the bottom of the culture hole, passes through the slit being fitted on the either side in hole
In protuberance alignment.Three dog bone hole positions are below the post so that tissue is formed around post.The end ratio of the mould
The length thereof of mould is deeper, and there is provided the hole of the agarose filled with fusing.Once the agarose is cooled to firm water-setting
Glue, post is pushed through agarose and entered in hole.Liquid cell/Collagen suspensions are pipetted into the hole, and organized
Formed at the top of agarose so that once tissue is removed from casting mould, the end of post is just exposed.In addition, in the light
The PDMS rings (or other types of ring) that fine end is formed can be used for preventing tissue from slipping.
G) exchange culture medium or add for test soluble compound (such as cardiac drug) usually require daily from
Bioreactor system is removed in incubator, and carefully replaces old culture medium with fresh culture using pipette.In order to keep away
Exempt from this troublesome process and ensure in the stable culture environment of the their entire life of the tissue, by fluid communication port
It is incorporated into the sidepiece of tissue-culture well.By the way that these ports are connected into culture medium source and waste media container by pipeline,
Culture medium is directly exchanged with the tissue for minimally destroying growth.Valve allow the system close when necessary and with stream
Body container is disconnected.Simple electric pump can be combined so that automatic change is cultivated in tissue bath (culture hole) as needed
Base.By increasing flow velocity, this can also provide mechanical shearing stress signal, or enhancing oxygen, nutrients and waste material to tissue
Transport.
H) (National Instruments are come from using the custom code write in LabVIEW programmed environments
Corp integrated, control) is carried out to whole system and is monitored.
The preferred embodiment of the system and equipment (it is intended to be illustrative and be not restrictive) has been described, notes
Meaning, can modify and change according to above-mentioned teaching those skilled in the art.It will thus be appreciated that can be in institute of the present invention
It is changed in disclosed specific embodiment, the scope and spirit of the present invention that these changes are summarized in such as appended claims
It is interior.
Claims (20)
1. bioreactor system, it configures the length control for being provided with engineering tissue and the one kind for measuring the engineering tissue
Or a variety of properties, including：
Housing comprising culture hole；
It is suspended at least one first flexible column in the culture hole；
At least one second rigid post in the culture hole is suspended at, second post is separated with first intercolumniation so that
The engineering tissue grows therebetween；
For the first equipment of the convergent force for measuring the engineering tissue, the equipment includes at least one first post, described
First post includes optical fiber, and the optical fiber is operatively coupled to light source and is configured to curved when the engineering tissue shrinks
Song, first equipment further includes the sensor for the mobile degree for being used to detect at least one first post；With
For the second equipment of the length for controlling the engineering tissue, second equipment includes at least one the second post of rigidity
And actuator, the actuator is operatively coupled at least one described second post, in a controlled manner in culture hole
At least one mobile described second post, so as to cause it is described between at least one first post and at least one described second post away from
From change, so as to control the length of engineering tissue.
2. bioreactor system according to claim 1, wherein the housing includes the pedestal for accommodating the culture hole
With removable lid, second equipment is fixedly attached to the lid.
3. bioreactor system according to claim 1, wherein three or more first posts and three or more
Second post is suspended in the culture hole, for growing three or more engineering tissue samples.
4. bioreactor system according to claim 3, wherein three or more described first post linear arrangements, with
And three or more described second post linear arrangements, at least one set of in face of post to limit, every group of post supports an engineering
Tissue samples so that three or more engineering tissue sample Parallel Growths.
5. bioreactor system according to claim 1, wherein first equipment includes pedestal, the pedestal couples
To the housing with least one first post described in positioning and hang in the culture hole.
6. bioreactor system according to claim 1, wherein first equipment is comprising being coupled to the optical fiber (the
One post) optical fiber LED emitter, the luminous point for producing the contact sensor, the sensor includes position sensing detection
Device, the position sensitive detectors are configured to measure the movement degree of optical fiber by measuring the position of luminous point to measure tissue receipts
Contracting.
7. bioreactor system according to claim 6, wherein the sole shape of the culture hole allows light to lead into material
The position sensitive detectors that road leads to below culture hole from the optical fiber.
8. bioreactor system according to claim 1, wherein the optical fiber has about 0.55mm diameter.
9. bioreactor system according to claim 2, wherein the optical fiber passes through the lid, close to the institute of the lid
The upper end for stating optical fiber represents the more rigid part of first post, and its underpart represents flexible portion, and flexible portion is received due to tissue
Contract in response to the power applied.
10. bioreactor system according to claim 1, wherein second post includes rigid tungsten post.
11. bioreactor system according to claim 1, wherein the actuator includes linear actuators, the line
Property actuator is operatively coupled to column base, and the column base is coupled at least one described second post.
12. bioreactor system according to claim 11, wherein the column base is operatively coupled to linear slide block,
The linear slide block causes column base linearly to move under the actuating of linear actuators, because one end of the engineering tissue is coupled
To the second post, the column base causes engineering tissue linearly to move.
13. bioreactor system according to claim 1, also comprising a pair of electrodes, the pair of electrode is arranged on institute
State in the culture hole outside at least one first post and at least one second post, for being carried across the length of the engineering tissue
Power supply is stimulated.
14. bioreactor system according to claim 13, wherein the electrode includes being arranged in the culture hole
Graphite plate electrode so that at least one described first post and at least one second post are disposed there between.
15. bioreactor system according to claim 1, wherein each in first and second equipment can be grasped
It is coupled to the controller for controlling its operation and function with making.
16. bioreactor system according to claim 1, also comprising for measuring the engineering tissue in the systole phase
Between the mechanism of action potential that produces.
17. bioreactor system according to claim 16, wherein the mechanism includes amplifier, the amplifier is matched somebody with somebody
It is equipped with so that its input is connected at least one described second post, enabling to the action electricity produced by the engineering tissue
Position carries out extracellular recording.
18. bioreactor system according to claim 1, wherein the culture hole includes built-in fluid communication end
Mouthful, the port allows medium and test agent to be fed into and flow out the culture hole.
19. bioreactor system according to claim 2, also comprising a pair of stimulating electrodes, the stimulating electrode is detachable
Ground is attached to the lid and is positioned in the culture hole so that at least one described first post and at least one second post are set
Put between the electrodes.
20. bioreactor system according to claim 19, wherein the electrode includes conductive magnet, the conductive magnetism
Body is used to electrode regularly but removably is attached into the lid so that electrode is maintained at fixed position, energy during use
It is enough easily to be removed from lid to be cleaned, and promptly replaced after its cleaning.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462046569P | 2014-09-05 | 2014-09-05 | |
| US62/046,569 | 2014-09-05 | ||
| PCT/US2015/046635 WO2016036532A1 (en) | 2014-09-05 | 2015-08-25 | Automated, multifunctional, engineered cardiac tissue culture and testing bioreactor system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107106605A true CN107106605A (en) | 2017-08-29 |
Family
ID=55440265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201580054406.0A Pending CN107106605A (en) | 2014-09-05 | 2015-08-25 | Automation, multi-functional, engineering heart tissue culture and test biology reactor assembly |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US10119109B2 (en) |
| EP (1) | EP3188739A4 (en) |
| CN (1) | CN107106605A (en) |
| WO (1) | WO2016036532A1 (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3176252A1 (en) * | 2015-12-04 | 2017-06-07 | Ludwig-Maximilians-Universität München | A system and method for determining a force applied to or generated by a cell or tissue culture |
| US10444064B2 (en) * | 2016-04-29 | 2019-10-15 | Optasense Holdings Limited | Monitoring a geological formation |
| US20190376014A1 (en) * | 2017-01-25 | 2019-12-12 | Igor R. Efimov | Apparatus and methods for in vitro preclinical human trials |
| US10656118B2 (en) * | 2017-02-02 | 2020-05-19 | Ika-Werke Gmbh & Co. Kg | Closure for an electrochemical vessel, electrochemical vessel and laboratory device |
| US11926844B2 (en) | 2017-08-15 | 2024-03-12 | Carnegie Mellon University | Three-dimensional microtissues with integrated mechanical loading |
| EP3717908A4 (en) * | 2017-11-29 | 2021-07-21 | Novoheart Limited | Bioreactor screening platform for modelling human systems biology and for screening for inotropic effects of agents on the heart |
| US20200199515A1 (en) * | 2018-02-01 | 2020-06-25 | Erin Glenn Roberts | Flexible device and its application for bio-cell in-vitro electrical and mechanical stimulation and characterization |
| WO2019186283A2 (en) * | 2018-03-28 | 2019-10-03 | Novoheart Limited | Modeling neurological disorders and ataxias with cardiac dysfunction using bioengineered heart tissues |
| CN111187719A (en) * | 2020-02-20 | 2020-05-22 | 西南交通大学 | Power-electricity coupling loading platform |
| US20230265373A1 (en) * | 2020-07-14 | 2023-08-24 | Novoheart International Limited | A Multi-Sample System for Engineered Tissue Strip Assays |
| NL2029002B1 (en) | 2021-08-19 | 2023-02-24 | Optics11 B V | Optical measurement of biological tissue. |
| CN113652349B (en) * | 2021-08-24 | 2023-07-07 | 威世药业（如皋）有限公司 | Development and preliminary pharmacokinetics experimental equipment for in-vivo analysis method of immediately-readjustment |
| EP4403621A1 (en) | 2023-01-19 | 2024-07-24 | Georg-August-Universität Göttingen Stiftung Öffentlichen Rechts | Device for and use of the device in cultivating tissue and observing contractions of the tissue |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090088680A1 (en) * | 2005-07-22 | 2009-04-02 | Alexander Aravanis | Optical tissue interface method and apparatus for stimulating cells |
| CN102859331A (en) * | 2010-04-30 | 2013-01-02 | 奥林巴斯株式会社 | Optical potentiometer and operation device |
| WO2014085933A1 (en) * | 2012-12-07 | 2014-06-12 | The Governing Council Of The University Of Toronto | Cardiac tissue constructs and methods of fabrication thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AUPQ578300A0 (en) * | 2000-02-22 | 2000-03-16 | Varian Australia Pty Ltd | Fluorescence measurement apparatus and method |
| US9493735B2 (en) * | 2005-04-15 | 2016-11-15 | Wake Forest University Health Sciences | Bioreactor system and method of enhancing functionality of muscle cultured in vitro |
| US20070281349A1 (en) * | 2006-06-06 | 2007-12-06 | West Virginia University | Industrial bioreactor and method of use in continuous protein and lipid recovery system |
| GB0613500D0 (en) * | 2006-07-07 | 2006-08-16 | Lectus Therapeutics Ltd | Apparatus and Methods |
| US20090105574A1 (en) * | 2007-10-23 | 2009-04-23 | National Yang-Ming University | Magnetic electrode device and an electrocardiograph detecting method thereof |
| US8172908B2 (en) | 2008-01-17 | 2012-05-08 | The University Of Hong Kong | Implant for tissue engineering |
| US8993346B2 (en) * | 2009-08-07 | 2015-03-31 | Nanomix, Inc. | Magnetic carbon nanotube based biodetection |
-
2015
- 2015-08-25 WO PCT/US2015/046635 patent/WO2016036532A1/en active Application Filing
- 2015-08-25 CN CN201580054406.0A patent/CN107106605A/en active Pending
- 2015-08-25 US US15/504,570 patent/US10119109B2/en active Active
- 2015-08-25 EP EP15837973.5A patent/EP3188739A4/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090088680A1 (en) * | 2005-07-22 | 2009-04-02 | Alexander Aravanis | Optical tissue interface method and apparatus for stimulating cells |
| CN102859331A (en) * | 2010-04-30 | 2013-01-02 | 奥林巴斯株式会社 | Optical potentiometer and operation device |
| WO2014085933A1 (en) * | 2012-12-07 | 2014-06-12 | The Governing Council Of The University Of Toronto | Cardiac tissue constructs and methods of fabrication thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016036532A1 (en) | 2016-03-10 |
| US10119109B2 (en) | 2018-11-06 |
| EP3188739A4 (en) | 2018-05-30 |
| US20170260488A1 (en) | 2017-09-14 |
| EP3188739A1 (en) | 2017-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107106605A (en) | Automation, multi-functional, engineering heart tissue culture and test biology reactor assembly | |
| Kook et al. | Neural probes for chronic applications | |
| US9206383B2 (en) | Bioreactor, devices, systems and methods | |
| Mousavi et al. | Measuring surface potential changes on leaves | |
| König et al. | A pulsatile bioreactor for conditioning of tissue-engineered cardiovascular constructs under endoscopic visualization | |
| KR20120121910A (en) | Integrated cultivation and measurement device for label-free detection and classification of cellular alterations, in particular for generation and characterisation of cell-spheroids, components and uses thereof | |
| Macdonald et al. | Recent advances in biomedical, biosensor and clinical measurement devices for use in humans and the potential application of these technologies for the study of physiology and disease in wild animals | |
| CN109486671A (en) | A kind of medical test microbial cultivation equipment | |
| Maschhoff et al. | An intelligent bioreactor system for the cultivation of a bioartificial vascular graft | |
| Staehlke et al. | Pulsed electrical stimulation affects osteoblast adhesion and calcium ion signaling | |
| CN110484442A (en) | Biological cell stimulating system and its control method based on annular spread electrode | |
| Rápalo et al. | Live cell imaging during mechanical stretch | |
| Castagnola et al. | Flexible glassy carbon multielectrode array for in vivo multisite detection of tonic and phasic dopamine concentrations | |
| Kunori et al. | An implantable cranial window using a collagen membrane for chronic voltage-sensitive dye imaging | |
| Ugolini et al. | Tailoring cardiac environment in microphysiological systems: an outlook on current and perspective heart-on-chip platforms | |
| Yallapragada et al. | ODX: A fitness tracker-based device for continuous bacterial growth monitoring | |
| Phillips et al. | Chlorella sensors in liquid marbles and droplets | |
| van Neste et al. | Designing a Bioreactor to Improve Data Acquisition and Model the Throughput of Engineered Cardiac Tissues | |
| Kusko et al. | Design, fabrication and characterization of a low-impedance 3D Electrode array system for neuro-electrophysiology | |
| CN208328035U (en) | Biological cell stimulating system based on annular spread electrode | |
| Yi et al. | 3D Printed Skull Cap and Benchtop Fabricated Microwire-Based Microelectrode Array for Custom Rat Brain Recordings | |
| Kim et al. | Fabrication of pillar shaped electrode arrays for artificial retinal implants | |
| Kim et al. | Investigating chorion softening of zebrafish embryos with a microrobotic force sensing system | |
| CN104007029A (en) | Dynamic mechanical experimental device and method for tissue engineering scaffold | |
| Pansodtee | Open-Hardware Electronic Embedded Systems for Biological Cell Culturing |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170829 |
|
| WD01 | Invention patent application deemed withdrawn after publication |